Molecular Cloning, Sequencing, and Expression of omp-40, the Gene Coding for the Major Outer Membrane Protein from the Acidophilic Bacterium Thiobacillus ferrooxidans

Thiobacillus ferrooxidans is one of the chemolithoautotrophic bacteria important in industrial biomining operations. Some of the surface components of this microorganism are probably involved in adaptation to their acidic environment and in bacterium-mineral interactions. We have isolated and characterized omp40, the gene coding for the major outer membrane protein from T. ferrooxidans. The deduced amino acid sequence of the Omp40 protein has 382 amino acids and a calculated molecular weight of 40,095.7. Omp40 forms an oligomeric structure of about 120 kDa that dissociates into the monomer (40 kDa) by heating in the presence of sodium dodecyl sulfate. The degree of identity of Omp40 amino acid sequence to porins from enterobacteria was only 22%. Nevertheless, multiple alignments of this sequence with those from several OmpC porins showed several important features conserved in the T. ferrooxidans surface protein, such as the approximate locations of 16 transmembrane beta strands, eight loops, including a large external L3 loop, and eight turns which allowed us to propose a putative 16-stranded beta-barrel porin structure for the protein. These results together with the previously known capacity of Omp40 to form ion channels in planar lipid bilayers strongly support its role as a porin in this chemolithoautotrophic acidophilic microorganism. Some characteristics of the Omp40 protein, such as the presence of a putative L3 loop with an estimated isoelectric point of 7.21 allow us to speculate that this can be the result of an adaptation of the acidophilic T. ferrooxidans to prevent free movement of protons across its outer membrane.     

Long-chain acyl CoA synthetase in microsomes from rat brain gray matter and white matter

Long-chain acyl coenzyme A (CoA) synthetase in homogenates and microsomes from rat brain gray and white matter was studied. The formation of the thioesters of CoA was studied upon addition of [1-¹⁴C]-labeled fatty acids. The maximal activities were seen with linoleic acid, followed by arachidonic, palmitic, and docosahexaenoic acids in both gray and white matter homogenates and microsomes. The specific activities in microsomes were 3–5 times higher than in homogenates. The presence of Triton X-100 in the assay system enhanced the activity of long-chain acyl CoA synthetase in homogenates. The effect was more pronounced in palmitic and docosahexaenoic acid activation. The apparentK _m values andV _max values for palmitic and docosahexaenoic acids were much lower than for linoleic and arachidonic acids. The presence of Triton X-100 in the medium caused a definite decrease in the apparentK _m and Vmax values for all the fatty acid except palmitic acid in which case the reverse was true. There were no significant differences observed in the kinetic measurements between gray and white matter microsomes. These findings are similar to those resulting from the known interference of Triton X-100 in the measurement of kinetic variables of long-chain acyl CoA synthetase of liver microsomes. In this work, no correlation was observed between the fatty acid composition of gray and white matter and the capacity of these tissues for the activation of different fatty acids.     

Isolation,Characterization and Functional Examination of the Gingival Immune Cell Network

Immune cell networks in tissues play a vital role in mediating local immunity and maintaining tissue homeostasis, yet little is known of the resident immune cell populations in the oral mucosa and gingiva. We have established a technique for the isolation and study of immune cells from murine gingival tissues, an area of constant microbial exposure and a vulnerable site to a common inflammatory disease, periodontitis. Our protocol allows for a detailed phenotypic characterization of the immune cell populations resident in the gingiva, even at steady state. Our procedure also yields sufficient cells with high viability for use in functional studies, such as the assessment of cytokine secretion ex vivo. This combination of phenotypic and functional characterization of the gingival immune cell network should aid towards investigating the mechanisms involved in oral immunity and periodontal homeostasis, but will also advance our understanding of the mechanisms involved in local immunopathology.     

Head-tail patterning of the vertebrate embryo: one, two or many unresolved problems?

When, where and how is the head-tail axis of the embryo set up during development? These are such fundamental and intensely studied questions that one might expect them to have been answered long ago. Not so; we still understand very little about the cellular or molecular mechanisms that lead to the orderly arrangement of body elements along the head-tail axis in vertebrates. In this paper, we outline some of the major outstanding problems and controversies and try to identify some reasons why it has been so difficult to resolve this important issue.     

Diel activity of adult pikeperch <Emphasis Type="Italic">Sander lucioperca</Emphasis> (L.) in a drainage canal in the Mediterranean basin during spring

Pikeperch (Sander lucioperca L.) is a broadly distributed fish species in Europe but little is known about its ecology in the southern part of its distribution area in warm climatic conditions. The aim of this study was to analyse pikeperch rate of movement and to assess whether it displayed a diel pattern related to temperature. Thus acoustic telemetry was used to track adult pikeperch in a drainage canal located in south of France. The survey was carried out in spring, during the spawning period. The results showed that females were more active than males. This is in accordance with previous data on the nest guardian behaviour of the males. For both genders, the activity rates increased during the study period as water temperature rose. Males and females displayed the same diel activity with a maximum at dusk, thereby confirming many indirect observations. Nevertheless, inter-individuals variations were observed. Thus, these results on diel activity are rather a general trend than a strict rule and suggest the involvement of other factors than light intensity in the control of diel activity. This diel rhythm is positively correlated to water temperature for females. Pikeperch activity may be the result of a trade-off between physiological requirements of temperature and light, satisfaction of energy needs and avoidance of predators.     

<Emphasis Type="Italic">In silico</Emphasis> coronary wave intensity analysis: application of an integrated one-dimensional and poromechanical model of cardiac perfusion

Coronary wave intensity analysis (cWIA) is a diagnostic technique based on invasive measurement of coronary pressure and velocity waveforms. The theory of WIA allows the forward- and backward-propagating coronary waves to be separated and attributed to their origin and timing, thus serving as a sensitive and specific cardiac functional indicator. In recent years, an increasing number of clinical studies have begun to establish associations between changes in specific waves and various diseases of myocardium and perfusion. These studies are, however, currently confined to a trial-and-error approach and are subject to technological limitations which may confound accurate interpretations. In this work, we have developed a biophysically based cardiac perfusion model which incorporates full ventricular–aortic–coronary coupling. This was achieved by integrating our previous work on one-dimensional modelling of vascular flow and poroelastic perfusion within an active myocardial mechanics framework. Extensive parameterisation was performed, yielding a close agreement with physiological levels of global coronary and myocardial function as well as experimentally observed cumulative wave intensity magnitudes. Results indicate a strong dependence of the backward suction wave on QRS duration and vascular resistance, the forward pushing wave on the rate of myocyte tension development, and the late forward pushing wave on the aortic valve dynamics. These findings are not only consistent with experimental observations, but offer a greater specificity to the wave-originating mechanisms, thus demonstrating the value of the integrated model as a tool for clinical investigation.     

The food web in the lower part of the Seine estuary: a synthesis of existing knowledge

The Seine estuary illustrates the alterations to estuaries due to human activities heavy releases of pollutants of various origins and significant morphological changes beginning in the middle of the 19th century. The intertidal mudflat surface has been seriously reduced (< 30km²) since the channels of the Seine River came under management. While the role of the Seine estuary in the dynamics of the eastern English Channel ecosystem is recognized as important, the biological characteristics of the estuary remained relatively unknown until the 1990s. Biological diversity was progressively impoverished from the polyhaline zone to the oligohaline zone. In spite of a heavily contaminated environment, the macrobenthic and planktonic fauna of the Seine estuary remains similar to those of other northeastern Atlantic estuaries. The fauna exhibit clear contrasts between areas with very high abundance and others with very low abundance. The pelagic fauna, especially the copepod Eurytemora affinisand the shrimp Palaemon longirostris, are more abundant in the Seine estuary than in other estuaries. Diversified and abundant, Abra alba-Pectinaria koreni and Macoma balthica benthic communities occur, respectively, in the outer and inner parts of the estuary. In subtidal flats, benthic fauna is especially poor in terms of specific richness, abundance and biomass. Paradoxically, considering the high abundance of prey, fish are particularly scarce. Two food webs have been identified. In the oligohaline zone, where turbidity is maximum, the food web is exclusively planktonic, due to dredging that prevented benthic fauna from settling. In the polyhaline zone, fish populations that feed particularly on benthic fauna benefit from low turbidity and high oxygen concentrations. So, in spite of heavy organic and metallic contamination and human activities, the Seine estuary remains a highly productive ecosystem, which provides a nursery for marine fish and feeding grounds for migratory birds. A global management plan appears to be necessary in order to guarantee that the Seine estuary continues to function as it currently does.     

Chemical Forms of Selenium in the Metal-Resistant Bacterium Ralstonia metallidurans CH34 Exposed to Selenite and Selenate

Ralstonia metallidurans CH34, a soil bacterium resistant to a variety of metals, is known to reduce selenite to intracellular granules of elemental selenium (Se⁰). We have studied the kinetics of selenite (Se^IV) and selenate (Se^VI) accumulation and used X-ray absorption spectroscopy to identify the accumulated form of selenate, as well as possible chemical intermediates during the transformation of these two oxyanions. When introduced during the lag phase, the presence of selenite increased the duration of this phase, as previously observed. Selenite introduction was followed by a period of slow uptake, during which the bacteria contained Se⁰ and alkyl selenide in equivalent proportions. This suggests that two reactions with similar kinetics take place: an assimilatory pathway leading to alkyl selenide and a slow detoxification pathway leading to Se⁰. Subsequently, selenite uptake strongly increased (up to 340 mg Se per g of proteins) and Se⁰ was the predominant transformation product, suggesting an activation of selenite transport and reduction systems after several hours of contact. Exposure to selenate did not induce an increase in the lag phase duration, and the bacteria accumulated approximately 25-fold less Se than when exposed to selenite. Se^IV was detected as a transient species in the first 12 h after selenate introduction, Se⁰ also occurred as a minor species, and the major accumulated form was alkyl selenide. Thus, in the present experimental conditions, selenate mostly follows an assimilatory pathway and the reduction pathway is not activated upon selenate exposure. These results show that R. metallidurans CH34 may be suitable for the remediation of selenite-, but not selenate-, contaminated environments.